毛竹C4H基因的鉴定及其表达模式分析

为了解毛竹（Phyllostachys edulis）中肉桂酸-4-羟化酶基因（C4H）的分子特征及其表达模式,采用生物信息学方法在毛竹基因组数据库中鉴定出6个C4H成员（PeC4H1~PeC4H6）,基因编码区长度为1 506~1 695 bp,推测编码501~564 aa,均具有保守的血红素结合域、苏氨酸结合槽基序和5个特征性底物识别位点,属于细胞色素P450超家族。系统进化分析表明,6个PeC4Hs可分为2类,分别含有2和4个成员。转录组数据分析表明,PeC4Hs在毛竹26个组织中的表达量存在明显差异,不同高度笋中PeC4Hs的表达差异显著。PeC4Hs启动子序列中含有多种响应逆境胁迫和激素信号的顺式调控元件,PeC4Hs表达受干旱和GA₃的影响,干旱时,仅PeC4H3/4在根中显著上调表达,其余成员均呈下调表达;GA₃处理下叶中PeC4H3/6迅速响应,呈先显著上调后逐渐降低的趋势,根中PeC4H2/5在处理前1 h短暂下调后又显著上调,至8 h时恢复到处理前的表达水平。因此,PeC4Hs可能在毛竹笋的木质化过程和应对非生物胁迫中发挥着重要作用。     

毛竹扩展蛋白基因的鉴定及其表达分析

为全面了解毛竹中扩展蛋白的分子特征和表达模式,本研究利用生物信息学方法在毛竹基因组中共鉴定出43个扩展蛋白基因家族成员,属于4个亚家族（EXPA、EXPB、EXLA和EXLB）,分别包含18、17、7和1个成员,分布在37个Scaffold上。除PeEXPA1没有内含子和PeEXLB1含有11个内含子外,其它毛竹扩展蛋白基因的内含子为1~5个。毛竹扩展蛋白基因编码蛋白长度为91~508个氨基酸,所有的氨基酸都具有高频密码子,大部分蛋白为碱性亲水性蛋白。大部分毛竹扩展蛋白二级结构中β转角占比例最少,而β折叠占比例最大,各亚家族多数成员具有类似的三级结构。qRT-PCR结果表明,18个EXPA亚家族成员在不同组织表达存在明显差异,除PeEXPA2、PeEXPA6外其它基因表达的最高值均出现在叶片中,表明它们可能在叶片生长过程中发挥着重要作用。     

毛竹SWEET基因家族的全基因组鉴定与分析

糖外排转运蛋白(Sugars will eventually be exported transporters, SWEETs)在植物生理活动和发育过程中起重要作用。为探究SWEET基因家族在毛竹(Phyllostachys edulis)的生长发育过程中起的作用,基于毛竹基因组数据,通过生物信息学方法对SWEET基因家族成员进行鉴定,并对其编码的蛋白质理化性质、系统进化及共线性关系、基因结构、启动子元件及表达模式、蛋白互作网路分析、GO注释等进行细致分析。研究结果表明:该家族基因结构、基序和结构域相对保守,所有成员均含有MtN3_slv结构域。上游启动子序列中含有多个同非生物胁迫以及生长发育相关元件,结合转录组表达量分析显示,多个家族成员在毛竹不同组织器官均有表达。共线性分析揭示毛竹SWEET家族在演化过程中存在全基因组多倍化事件。蛋白互作网路分析挖掘出2个重要核心家族成员,GO注释分析进一步证实毛竹SWEET主要负责体内糖类物质的转运。以上结果为毛竹SWEET基因功能鉴定提供了重要参考,对于毛竹快速生长分子机制研究打下了基础。     

Comprehensive Analysis of Wall-Associated Kinase Genes and Their Expression Under Abiotic and Biotic Stress in Chinese Cabbage (Brassica rapa ssp. pekinensis)

The wall-associated kinase (WAK) gene family, a subfamily of the receptor-like kinase (RLK) gene family, is associated with the cell wall in plants, and has vital functions in cell expansion, pathogen resistance, and heavy metal stress tolerance because of their roles of the extracellular environment sensors to trigger intracellular signals in Arabidopsis. In the present study, 96 Chinese cabbage (Brassica rapa ssp. pekinensis) BrWAK gene family members were identified from the B. rapa genome using a reiterative database search and manual confirmation. The protein domain characterization, gene structure analysis, and phylogenetic analysis of the BrWAKs classified them into three gene groups. Comparative genomic analysis between WAK genes from Chinese cabbage and Arabidopsis revealed that the BrWAK genes have undergone the gene expansion and deletion events during evolution. Furthermore, the conserved motifs in the kinase domains of the WAK proteins and eukaryotic protein kinase family proteins were compared and some non-RD kinase proteins among the BrWAKs were identified. Ultimately, expression analysis of BrWAK genes in six tissues and under various stress conditions revealed that some tissue-specific WAK genes might function in callus cell growth and reproduction process; Bra012273, Bra016426, Bra016427, and Bra025882 might be involved in downy mildew resistance and high humidity stress; Bra012273, Bra025882, and Bra025883 might be responded to drought and heat stress. Taken together, this research was identified and classified the WAK gene family in Chinese cabbage and provided valuable resources to explore the potential roles of BrWAK genes in plant development and stress responses.

     

Genome-Wide Identification of the F-box Gene Family and Expression Analysis under Drought and Salt Stress in Barley

The F-box protein-encoding gene family plays an essential role in plant stress resistance. In present study, 126 non-redundant F-box genes were identified in barley (Hordeum vulgare L., Hv). The corresponding proteins contained 165– 887 amino acid residues and all were amphiphilic, except 5 proteins. Phylogenetic analysis of F-box protein sequences in barley and stress-related F-box protein sequences in wheat and Arabidopsis thaliana (At) was used to classify barley F-box genes are divided into 9 subfamilies (A–I). A structure-based sequence alignment demonstrated that F-box proteins were highly conserved with a total of 10 conserved motifs. In total, 124 F-box genes were unevenly distributed on 7 chromosomes; another 2 genes have not been anchored yet. The gene structure analysis revealed high variability in the number of exons and introns in F-box genes. Comprehensive analysis of expression profiles and phylogenetic tree analysis, a total of 12 F-box genes that may be related to stress tolerance in barley were screened. Of the 12 detected F-box genes, 8 and 10 were upregulated after drought and salt stress treatments, respectively, using quantitative real-time polymerase chain reaction (qRT-PCR). This study is the first systematic analysis conducted on the F-box gene family in barley, which is of great importance for clarifying this family’s bioinformatic characteristics and elucidating its function in barley stress resistance. These results will serve as a theoretical reference for subsequent research on molecular regulation mechanisms, genetic breeding, and improvement.     

漓江上游毛竹生理生态特征对不同土壤水分的响应

漓江上游毛竹林面积大,是我国毛竹最南端产区。该研究针对当地季节性干旱问题,通过模拟降水使土壤水分变化,共设立了5个毛竹水分处理小区(CK.对照;A.无降水+覆膜;B.降水5 mm+覆膜;C.降水10mm+覆膜;D.降水20 mm+覆膜),并与HM(木荷Schima superba)(自然状态)进行对比。结果表明:土壤水分变化影响了毛竹叶片水势和叶绿素的变化,叶片白天水势下降,傍晚均可恢复到凌晨水势。C处理的毛竹午间水势下降值最小,叶绿素含量也最高。本研究区位于最南端产区,毛竹的光合生产力也属偏低水平。适当的土壤水分亏缺,毛竹表现出相对的高净光合速率(P_(n))、高蒸腾速率(Tr)、低水分利用效率(WUE)的特点;过多或过少土壤水分,则为低P_(n)和Tr,但高WUE。毛竹叶片的P_(n)与气孔导度Gs呈极显著的正相关关系,说明毛竹的光合速率受气孔调节明显;Tr与午时叶水势呈负相关(符合二项式函数)关系,土壤水分问题造成的叶片水分不足同时也影响了毛竹的Tr。水分亏缺,P_(n)主要由气孔调节,但水分过多导致P_(n)的下降应该是由气孔导度的下降和叶肉细胞光合能力的下降共同作用的结果。水分过少或过多均对毛竹生理生态过程产生负效应。相对于木荷,毛竹的P_(n)较高,但同时也消耗更多的水分。     

毛竹油菜素内酯受体激酶基因的分子特征及表达模式分析

为探究毛竹(Phyllostachys edulis)油菜素内酯(brassinolide,BL)受体激酶基因的分子特征和表达模式,采用生物信息学方法对毛竹中BL受体激酶基因进行了分析,并应用实时定量PCR技术对基因的表达模式进行了研究。结果表明,在毛竹基因组中共获得8条BL受体激酶基因同源序列(PeBRLs),分别属于4个亚家族。8个PeBRLs编码858~1 224氨基酸,分子量为92~130 kDa。PeBRLs结构相对保守,激酶区均具有BL受体激酶特有的3个保守结构域;除PeBRL1-1具有2个跨膜结构域外,其余PeBRLs只有1个跨膜结构域。8个PeBRLs全部定位在细胞膜上,属于典型的膜嵌合蛋白。实时定量PCR结果显示,每个亚家族成员基因的组织特异性表达模式基本一致,但不同亚家族之间差异明显;在不同发育阶段的竹笋中,PeBRLs的表达呈现为4种变化趋势。因此,8个PeBRLs在毛竹不同组织和笋的不同发育阶段可能发挥着不同的作用。     

Contrasting duplication patterns reflect functional diversities of ubiquitin and ubiquitin‐like protein modifiers in plants

Ubiquitin (Ub) and Ub‐like proteins, collectively forming the ubiquiton family, regulate nearly all aspects of cellular processes via post‐translational modifications. Studies devoted to specific members suggested a large expansion of this family in plants; however, a lack of systematic analysis hinders the comparison of individual members at both evolutionary history and functional divergence levels, which may provide new insight into biological functions. In this work, we first retrieved a total of 5856 members of 17 known ubiquiton subfamilies in 50 plant genomes by searching both prior annotations and missing loci in each genome. We then applied this list to analyze the duplication history of major ubiquiton subfamilies in plants. We show that autophagy‐related protein 8 (ATG8), membrane‐anchored Ub‐fold (MUB), small Ub‐like modifier (SUMO) and Ub loci encode 88% of the plant ubiquiton family. Although whole genome duplications (WGDs) significantly expanded the family, we discovered contrasting duplication patterns both in species and in subfamilies. Within the family, the ATG8 and MUB members were primarily duplicated through WGDs, whereas a significant number of Ub and SUMO loci were generated through retroposition and tandem duplications, respectively. Although Ub coding regions are highly conserved in plants, promoter activity analysis demonstrated lineage‐specific expression patterns of polyUb genes in Oryza sativa (rice) and Arabidopsis, confirming their retroposition origin. Based on the theory of dosage balance constraints, our study suggests that ubiquiton members duplicated through WGDs play crucial roles in plants, and that the regulatory pathways involving ATG8 and MUB are more conserved than those controlled by Ub and SUMO.     

苹果SRS基因家族的鉴定与功能分析北大核心CSCD

SHI-related sequence(SRS)基因家族通过介导激素变化以调控植物成花及生长发育,并且在适应环境胁迫中起重要调控作用。该研究基于苹果(Malus domestica Borkh.)基因组数据,通过生物信息学手段鉴定苹果SRS基因家族成员,并分析SRS基因家族特点与功能及表达情况。结果表明:(1)苹果MdSRS基因家族共包含11个成员,分别命名为MdSRS1-MdSRS11,不均匀地分布在苹果的9条染色体上。(2)MdSRS蛋白包含229~414个不等的氨基酸残基,等电点分布在6.38~9.36之间;亚细胞定位结果表明,MdSRS蛋白大多分布于细胞膜,在细胞核、叶绿体中也有分布。(3)通过引入拟南芥、水稻、番茄及杨树的SRS基因进行系统发育分析表明,将11个MdSRSs分成5个亚族(A-A),在A4中分布最多。(4)顺式作用元件分析表明,11个MdSRSs启动子上游2 000 bp序列分布有激素、环境适应性和逆境诱导等响应元件。(5)荧光定量PCR结果显示,苹果MdSRS基因家族在盐胁迫和干旱胁迫下总体呈下调表达,在ABA胁迫后大多呈上调表达,是具有很大潜力的抗性候选基因,说明SRS家族对ABA调节等非生物胁迫具有调控作用。研究认为,SRS家族的11个成员均参与了调控干旱、盐及ABA胁迫多种逆境的响应,推测在实际苹果生产中对抵御不良环境具有重要作用。     

Genome size and sequence composition of moso bamboo: A comparative study

Moso bamboo (Phyllostachys pubescens) is one of the world’s most important bamboo species. It has the largest area of all planted bamboo—over two-thirds of the total bamboo forest area—and the highest economic value in China. Moso bamboo is a tetraploid (4x=48) and a special member of the grasses family. Although several genomes have been sequenced or are being sequenced in the grasses family, we know little about the genome of the bambusoids (bamboos). In this study, the moso bamboo genome size was estimated to be about 2034 Mb by flow cytometry (FCM), using maize (cv. B73) and rice (cv. Nipponbare) as internal references. The rice genome has been sequenced and the maize genome is being sequenced. We found that the size of the moso bamboo genome was similar to that of maize but significantly larger than that of rice. To determine whether the bamboo genome had a high proportion of repeat elements, similar to that of the maize genome, approximately 1000 genome survey sequences (GSS) were generated. Sequence analysis showed that the proportion of repeat elements was 23.3% for the bamboo genome, which is significantly lower than that of the maize genome (65.7%). The bamboo repeat elements were mainly Gypsy/DIRS1 and Ty1/Copia LTR retrotransposons (14.7%), with a few DNA transposons. However, more genomic sequences are needed to confirm the above results due to several factors, such as the limitation of our GSS data. This study is the first to investigate sequence composition of the bamboo genome. Our results are valuable for future genome research of moso and other bamboos.